A metabolomic perspective of pazopanib-induced acute hepatotoxicity in mice

To elucidate the metabolism of pazopanib, a metabolomics approach was performed based on ultra-performance liquid chromatography coupled with electrospray ionization quadrupole mass spectrometry.A total of 22 pazopanib metabolites were identified <i>in vitro</i> and <i>in vivo</i>. Among these metabolites, 17 were novel, including several cysteine adducts and aldehyde derivatives. By screening using recombinant CYPs, CYP3A4 and CYP1A2 were found to be the main forms involved in the pazopanib hydroxylation. Formation of a cysteine conjugate (<b>M3</b>), an aldehyde derivative (<b>M15</b>) and two <i>N</i>-oxide metabolites (<b>M18</b> and <b>M20</b>) from pazopanib could induce the oxidative stress that may be responsible in part for pazopanib-induced hepatotoxicity.Morphological observation of the liver suggested that pazopanib (300 mg/kg) could cause liver injury. The aspartate transaminase and alanine aminotransferase in serum significantly increased after pazopanib (150, 300 mg/kg) treatment; this liver injury could be partially reversed by the broad-spectrum CYP inhibitor 1-aminobenzotriazole (ABT). Metabolomics analysis revealed that pazopanib could significantly change the levels of <i>L</i>-carnitine, proline and lysophosphatidylcholine 18:1 in liver. Additionally, drug metabolism-related gene expression analysis revealed that hepatic <i>Cyp2d22</i> and <i>Abcb1a</i> (P-gp) mRNAs were significantly lowered by pazopanib treatment.In conclusion, this study provides a global view of pazopanib metabolism and clues to its influence on hepatic function. To elucidate the metabolism of pazopanib, a metabolomics approach was performed based on ultra-performance liquid chromatography coupled with electrospray ionization quadrupole mass spectrometry. A total of 22 pazopanib metabolites were identified <i>in vitro</i> and <i>in vivo</i>. Among these metabolites, 17 were novel, including several cysteine adducts and aldehyde derivatives. By screening using recombinant CYPs, CYP3A4 and CYP1A2 were found to be the main forms involved in the pazopanib hydroxylation. Formation of a cysteine conjugate (<b>M3</b>), an aldehyde derivative (<b>M15</b>) and two <i>N</i>-oxide metabolites (<b>M18</b> and <b>M20</b>) from pazopanib could induce the oxidative stress that may be responsible in part for pazopanib-induced hepatotoxicity. Morphological observation of the liver suggested that pazopanib (300 mg/kg) could cause liver injury. The aspartate transaminase and alanine aminotransferase in serum significantly increased after pazopanib (150, 300 mg/kg) treatment; this liver injury could be partially reversed by the broad-spectrum CYP inhibitor 1-aminobenzotriazole (ABT). Metabolomics analysis revealed that pazopanib could significantly change the levels of <i>L</i>-carnitine, proline and lysophosphatidylcholine 18:1 in liver. Additionally, drug metabolism-related gene expression analysis revealed that hepatic <i>Cyp2d22</i> and <i>Abcb1a</i> (P-gp) mRNAs were significantly lowered by pazopanib treatment. In conclusion, this study provides a global view of pazopanib metabolism and clues to its influence on hepatic function.